RNA Interference – Natural Gene-Based Technology for Highly Specific Pest Control (HiSPeC)

RNAi technologies are more environmentally friendly, as the technology provides greater specificity in pest targeting, while reducing the potential negative effects on ecosystems and leaving beneficial insects and other organisms unharmed in crop ecosystems. Consequently, the increase in native fauna improves the efficacy of biological control agents against pests and pathogens. A growing understanding of the ubiquitous nature of RNAi, along with evidence for efficient, non-transgenic, topical applications has already begun to garner support among organic and industry producers. Designing solutions to agricultural problems based upon the same mechanisms used in nature provides newer, safer solutions to pests and pathogens for all agricultural industries

SOLENOPSIS INVICTA VIRUS (SINV-1) INFECTION AND INSECTICIDE INTERACTIONS IN THE RED IMPORTED FIRE ANT (HYMENOPTERA: FORMICIDAE)

Controlling invasive species is a growing concern; however, pesticides can be detrimental for non-target organisms. The red imported fire ant (Solenopsis invicta Buren; Hymenoptera: Formicidae) has aggressively invaded ~138 million ha in the USA and causes over $6 billion in damage and control efforts annually (Valles 2011). Myriad research studies have been conducted to discover safe biological control agents to manage these invasive pests (Valles et al. 2004; Milks et al. 2008; Oi et al. 2009; Yang et al. 2009; Wang et al. 2010; Callcott et al. 2011; Porter et al. 2011; Tufts et al. 2011). Viruses may be lethal due to modifications of cellular processes and induction of defense responses or may produce distinct survival outcomes depending on species (i.e. ascoviruses) (Stasiak et al. 2005). The Solenopsis invicta virus (SINV-1) is a positive sense, single-stranded RNA virus, which can only infect the genus Solenopsis at all stages of development, and is verticallytransmitted within a colony (Valles et al. 2004; Valles 2012)

Asian citrus psyllid RNAi pathway : RNAi evidence

Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if the RNAi machinery could be exploited to develop a management strategy for this pest. Analyses of RNAi-related genes in the Asian citrus psyllid genome showed an absence of sequences encoding R2D2, a dsRNA-binding protein that functions as a cofactor of Dicer-2 in Drosophila. Nevertheless, bioassays using an in Planta System showed that the Asian citrus psyllid was very sensitive to ingested dsRNA, demonstrating a strong RNAi response. A small dose of dsRNA administered through a citrus flush was enough to trigger the RNAi mechanism, causing significant suppression of the targeted transcript, and increased psyllid mortality. This study provides evidence of a functional RNAi machinery, which could be further exploited to develop RNAi based management strategies for the control of the Asian citrus psyllid

Analysis and Functional Annotation of Expressed Sequence Tags from the Asian Longhorned Beetle, Anoplophora glabripennis

The Asian longhorned beetle, Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae), is one of the most economically and ecologically devastating forest insects to invade North America in recent years. Despite its substantial impact, limited effort has been expended to define the genetic and molecular make-up of this species. Considering the significant role played by late-stadia larvae in host tree decimation, a small-scale EST sequencing project was done using a cDNA library constructed from 5th -instar A. glabripennis. The resultant dataset consisted of 599 high quality ESTs that, upon assembly, yielded 381 potentially unique transcripts. Each of these transcripts was catalogued as to putative molecular function, biological process, and associated cellular component according to the Gene Ontology classification system. Using this annotated dataset, a subset of assembled sequences was identified that are putatively associated with A. glabnpennis development and metamorphosis. This work will contribute to understanding of the diverse molecular mechanisms that underlie coleopteran morphogenesis and enable the future development of novel control strategies for management of this insect pest

Working Together for Mental Health: Evaluation of a one-day mental health course for human service providers

BACKGROUND: The Working Together For Mental Health course is an 8-hour course designed to demystify mental illness and mental health services. The main target group for the course is people working in human service organisations who provide services for people with mental illness. METHODS: A questionnaire was administered to all participants attending the course during 2003 (n = 165). Participants completed the questionnaire before and immediately after the course, and at three month follow-up. RESULTS: A response rate of 69% was achieved with 114 people completing the questionnaire on all three occasions. The responses showed a significant improvement in the self-assessed knowledge and confidence of participants to provide human services to people with a mental health problem or disorder, three months after the course. There was no significant improvement in participants' attitudes or beliefs about people with a mental health problem or disorder at three month follow-up; however, participants' attitudes were largely positive before entering the course. CONCLUSION: The Working Together For Mental Health course was successful in improving participants' confidence and knowledge around providing human services to people with a mental health illness

Gene content evolution in the arthropods

Arthropods comprise the largest and most diverse phylum on Earth and play vital roles in nearly every ecosystem. Their diversity stems in part from variations on a conserved body plan, resulting from and recorded in adaptive changes in the genome. Dissection of the genomic record of sequence change enables broad questions regarding genome evolution to be addressed, even across hyper-diverse taxa within arthropods. Using 76 whole genome sequences representing 21 orders spanning more than 500 million years of arthropod evolution, we document changes in gene and protein domain content and provide temporal and phylogenetic context for interpreting these innovations. We identify many novel gene families that arose early in the evolution of arthropods and during the diversification of insects into modern orders. We reveal unexpected variation in patterns of DNA methylation across arthropods and examples of gene family and protein domain evolution coincident with the appearance of notable phenotypic and physiological adaptations such as flight, metamorphosis, sociality, and chemoperception. These analyses demonstrate how large-scale comparative genomics can provide broad new insights into the genotype to phenotype map and generate testable hypotheses about the evolution of animal diversity

Non-Compliance with Growth Hormone Treatment in Children Is Common and Impairs Linear Growth

BACKGROUND: GH therapy requires daily injections over many years and compliance can be difficult to sustain. As growth hormone (GH) is expensive, non-compliance is likely to lead to suboptimal growth, at considerable cost. Thus, we aimed to assess the compliance rate of children and adolescents with GH treatment in New Zealand. METHODS: This was a national survey of GH compliance, in which all children receiving government-funded GH for a four-month interval were included. Compliance was defined as ≥ 85% adherence (no more than one missed dose a week on average) to prescribed treatment. Compliance was determined based on two parameters: either the number of GH vials requested (GHreq) by the family or the number of empty GH vials returned (GHret). Data are presented as mean ± SEM. FINDINGS: 177 patients were receiving GH in the study period, aged 12.1 ± 0.6 years. The rate of returned vials, but not number of vials requested, was positively associated with HVSDS (p < 0.05), such that patients with good compliance had significantly greater linear growth over the study period (p<0.05). GHret was therefore used for subsequent analyses. 66% of patients were non-compliant, and this outcome was not affected by sex, age or clinical diagnosis. However, Maori ethnicity was associated with a lower rate of compliance. INTERPRETATION: An objective assessment of compliance such as returned vials is much more reliable than compliance based on parental or patient based information. Non-compliance with GH treatment is common, and associated with reduced linear growth. Non-compliance should be considered in all patients with apparently suboptimal response to GH treatment

Sensitive and Specific Fluorescent Probes for Functional Analysis of the Three Major Types of Mammalian ABC Transporters

An underlying mechanism for multi drug resistance (MDR) is up-regulation of the transmembrane ATP-binding cassette (ABC) transporter proteins. ABC transporters also determine the general fate and effect of pharmaceutical agents in the body. The three major types of ABC transporters are MDR1 (P-gp, P-glycoprotein, ABCB1), MRP1/2 (ABCC1/2) and BCRP/MXR (ABCG2) proteins. Flow cytometry (FCM) allows determination of the functional expression levels of ABC transporters in live cells, but most dyes used as indicators (rhodamine 123, DiOC2(3), calcein-AM) have limited applicability as they do not detect all three major types of ABC transporters. Dyes with broad coverage (such as doxorubicin, daunorubicin and mitoxantrone) lack sensitivity due to overall dimness and thus may yield a significant percentage of false negative results. We describe two novel fluorescent probes that are substrates for all three common types of ABC transporters and can serve as indicators of MDR in flow cytometry assays using live cells. The probes exhibit fast internalization, favorable uptake/efflux kinetics and high sensitivity of MDR detection, as established by multidrug resistance activity factor (MAF) values and Kolmogorov-Smirnov statistical analysis. Used in combination with general or specific inhibitors of ABC transporters, both dyes readily identify functional efflux and are capable of detecting small levels of efflux as well as defining the type of multidrug resistance. The assay can be applied to the screening of putative modulators of ABC transporters, facilitating rapid, reproducible, specific and relatively simple functional detection of ABC transporter activity, and ready implementation on widely available instruments

Structure-based analysis of CysZ-mediated cellular uptake of sulfate

Sulfur, most abundantly found in the environment as sulfate (SO42-), is an essential element in metabolites required by all living cells, including amino acids, co-factors and vitamins. However, current understanding of the cellular delivery of SO42- at the molecular level is limited. CysZ has been described as a SO42- permease, but its sequence family is without known structural precedent. Based on crystallographic structure information, SO42- binding and flux experiments, we provide insight into the molecular mechanism of CysZ-mediated translocation of SO42- across membranes. CysZ structures from three different bacterial species display a hitherto unknown fold and have subunits organized with inverted transmembrane topology. CysZ from Pseudomonas denitrificans assembles as a trimer of antiparallel dimers and the CysZ structures from two other species recapitulate dimers from this assembly. Mutational studies highlight the functional relevance of conserved CysZ residues

Micro-CT study of male genitalia and reproductive system of the Asian citrus psyllid, Diaphorina citri Kuwayama, 1908 (Insecta: Hemiptera, Liviidae)

The Asian citrus psyllid (ACP), Diaphorina citri, is a major vector of the bacteria Candidatus Liberibacter asiaticus and C.L. americanus, which cause Huanglongbing disease (HLB) (aka Citrus greening disease), considered the most serious bacterial disease of citrus trees. As part of a multidisciplinary project on psyllid biology (www.citrusgreening.org), the results presented here concern a detailed anatomical study of the male reproductive system (testes, seminal vesicles, accessory glands, sperm pump, connecting ducts, and aedeagus) using micro-computed tomography (micro-CT). The study summarizes current knowledge on psyllids male reproductive system and represents significant advances in the knowledge of ACP anatomy.This work was supported by USDA-NIFA Award 2014-70016-23028 ªDeveloping an Infrastructure and Product Test Pipeline to Deliver Novel Therapies for Citrus Greening Diseaseº, 2015-2020